Generation of a tightly regulated doxycycline-inducible model for studying mouse intestinal biology.
To develop a sensitive and inducible system to study intestinal biology, we generated a transgenic mouse model expressing the reverse tetracycline transactivator rtTA2-M2 under control of the 12.4 kb murine Villin promoter. The newly generated Villin-rtTA2-M2 mice were then bred with the previously developed tetO-HIST1H2BJ/GFP model to assess inducibility and tissue-specificity. Expression of the histone H2B-GFP fusion protein was observed exclusively upon doxycycline induction and was uniformly distributed throughout the intestinal epithelium. The Villin-rtTA2-M2 was also found to drive transgene expression in the developing mouse intestine. Furthermore, we could detect transgene expression in the proximal tubules of the kidney and in a population of alleged gastric progenitor cells. By administering different concentrations of doxycycline, we show that the Villin-rtTA2-M2 system drives transgene expression in a dosage-dependent fashion. Thus, we have generated a novel doxycycline-inducible mouse model, providing a valuable tool to study the effect of different gene dosages on intestinal physiology and pathology.
Roth
Sabrina
S
Department of Pathology, Josephine Nefkens Institute, Erasmus MC, Rotterdam, The Netherlands.
Franken
Patrick
P
van Veelen
Wendy
W
Blonden
Lau
L
Raghoebir
Lalini
L
Beverloo
Berna
B
van Drunen
Ellen
E
Kuipers
Ernst J
EJ
Rottier
Robbert
R
Fodde
Riccardo
R
Smits
Ron
R
D000818:Animals; D004318:Doxycycline / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects*; D017930:Genes, Reporter / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D009928:Organ Specificity; D012106:Research; D017186:Titrimetry; D019076:Transgenes / Q000235:genetics
D004318:Doxycycline